Cloned (Comment) | Organism |
---|---|
gene GDH1, DNA and amino acid sequence determination and analysis, overexpression as His6-tagged protein in Escherichia coli strain KRX | Plasmodium falciparum |
gene GDH1, expression as His6-tagged enzyme in Escherichia coli strain M15, method optimization | Plasmodium falciparum |
Crystallization (Comment) | Organism |
---|---|
purified recombinant detagged GDH2, 0.0025 ml of 16 mg/ml protein in 100 mM Tris, 500 mM NaCl, pH 7.8, mixed with 0.0025 ml of reservoir solution containing 0.2 M magnesium chloride hexahydrate, 0.1 M Tris/HCl pH 8.5, 30% PEG 4000 including 0.01 M spermine tetra HCl droplet concentration and 10 mM glutamate, X-ray diffraction structure determination and analysis at 3.1 A resolution | Plasmodium falciparum |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
bithionol | - |
Plasmodium falciparum | |
GW5074 | - |
Plasmodium falciparum | |
Hexachlorophene | 15% inhibition at 0.016 mM; 80% inhibition at 0.016 mM | Plasmodium falciparum | |
additional information | inhibitors hexachlorophene, GW5074, and bithionol are more effective on PfGDH2 than on PfGDH1; inhibitors hexachlorophene, GW5074, and bithionol are more effective on PfGDH2 than on PfGDH1 | Plasmodium falciparum |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.007 | - |
NADPH | pH 7.0, 25°C, recombinant enzyme | Plasmodium falciparum | |
0.012 | - |
NADPH | pH 7.0, 25°C, recombinant enzyme | Plasmodium falciparum | |
0.019 | - |
NADP+ | pH 8.0, 25°C, recombinant enzyme | Plasmodium falciparum | |
0.02 | - |
NADP+ | pH 8.0, 25°C, recombinant enzyme | Plasmodium falciparum | |
0.2 | - |
2-oxoglutarate | pH 7.0, 25°C, recombinant enzyme | Plasmodium falciparum | |
0.365 | - |
2-oxoglutarate | pH 7.0, 25°C, recombinant enzyme | Plasmodium falciparum | |
0.61 | - |
L-glutamate | pH 8.0, 25°C, recombinant enzyme | Plasmodium falciparum | |
1.05 | - |
L-glutamate | pH 8.0, 25°C, recombinant enzyme | Plasmodium falciparum |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
apicoplast | GDH2 co-localizes with the acyl carrier protein | Plasmodium falciparum | 20011 | - |
cytosol | - |
Plasmodium falciparum | 5829 | - |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
52546 | - |
6 * 52546, GDH1, sequence calculation | Plasmodium falciparum |
54024 | - |
6 * 54024, GDH2, sequence calculation | Plasmodium falciparum |
300000 | - |
recombinant enzyme, gel filtration | Plasmodium falciparum |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
2-oxoglutarate + NH3 + NADPH + H+ | Plasmodium falciparum | - |
L-glutamate + H2O + NADP+ | - |
r | |
L-glutamate + H2O + NADP+ | Plasmodium falciparum | - |
2-oxoglutarate + NH3 + NADPH + H+ | - |
r | |
additional information | Plasmodium falciparum | Like PfGDH1, PfGDH2 is an NADP(H)-dependent enzyme with a specific activity comparable to PfGDH1 but with slightly higher Km values for its substrates | ? | - |
? | |
additional information | Plasmodium falciparum | PfGDH1, like PfGDH2, is an NADP(H)-dependent enzyme with a specific activity comparable to PfGDH2 but with slightly lower Km values for its substrates | ? | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Plasmodium falciparum | Q8ILF7 | GDH2 | - |
Plasmodium falciparum | Q8ILT0 | GDH1 | - |
Purification (Comment) | Organism |
---|---|
recombinant His6-tagged GDH2 from Escherichia coli strain KRX by nickel affinity chromatography and gel filtration | Plasmodium falciparum |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
4 | - |
deamination, pH 8.0, 25°C, recombinant enzyme | Plasmodium falciparum |
9 | - |
deamination, pH 8.0, 25°C, recombinant enzyme | Plasmodium falciparum |
28 | - |
amination, pH 8.0, 25°C, recombinant enzyme | Plasmodium falciparum |
61 | - |
amination, pH 8.0, 25°C, recombinant enzyme | Plasmodium falciparum |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
2-oxoglutarate + NH3 + NADPH + H+ | - |
Plasmodium falciparum | L-glutamate + H2O + NADP+ | - |
r | |
2-oxoglutarate + NH3 + NADPH + H+ | referred reaction | Plasmodium falciparum | L-glutamate + H2O + NADP+ | - |
r | |
L-glutamate + H2O + NADP+ | - |
Plasmodium falciparum | 2-oxoglutarate + NH3 + NADPH + H+ | - |
r | |
additional information | Like PfGDH1, PfGDH2 is an NADP(H)-dependent enzyme with a specific activity comparable to PfGDH1 but with slightly higher Km values for its substrates | Plasmodium falciparum | ? | - |
? | |
additional information | PfGDH1, like PfGDH2, is an NADP(H)-dependent enzyme with a specific activity comparable to PfGDH2 but with slightly lower Km values for its substrates | Plasmodium falciparum | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
hexamer | 6 * 52546, GDH1, sequence calculation | Plasmodium falciparum |
hexamer | 6 * 54024, GDH2, sequence calculation | Plasmodium falciparum |
More | the subunit interactions are mediated by networks of salt bridges and hydrogen bonds, monome-monomer interaction, overview | Plasmodium falciparum |
More | the three-dimensional structure of hexameric PfGDH2 is solved to 3.1 A resolution, overview. The subunit interactions are mainly assisted by hydrogen bonds and hydrophobic interactions, monome-monomer interaction, overview | Plasmodium falciparum |
Synonyms | Comment | Organism |
---|---|---|
GDH1 | - |
Plasmodium falciparum |
GDH2 | - |
Plasmodium falciparum |
glutamate dehydrogenase 1 | - |
Plasmodium falciparum |
glutamate dehydrogenase 2 | - |
Plasmodium falciparum |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Plasmodium falciparum |
25 | - |
amination, assay at | Plasmodium falciparum |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
80 | - |
purified enzyme, 10 min, stable | Plasmodium falciparum |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.367 | - |
L-glutamate | pH 8.0, 25°C, recombinant enzyme | Plasmodium falciparum | |
25 | - |
2-oxoglutarate | pH 7.0, 25°C, recombinant enzyme | Plasmodium falciparum | |
53.33 | - |
2-oxoglutarate | pH 7.0, 25°C, recombinant enzyme | Plasmodium falciparum | |
78.33 | - |
L-glutamate | pH 8.0, 25°C, recombinant enzyme | Plasmodium falciparum |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7 | - |
amination, assay at | Plasmodium falciparum |
8 | - |
deamination, assay at | Plasmodium falciparum |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
additional information | no activity with NAD+ | Plasmodium falciparum | |
NADP+ | - |
Plasmodium falciparum | |
NADPH | - |
Plasmodium falciparum |
Organism | Comment | pI Value Maximum | pI Value |
---|---|---|---|
Plasmodium falciparum | sequence calculation | - |
7.48 |
Plasmodium falciparum | sequence calculation | - |
9.09 |
IC50 Value | IC50 Value Maximum | Comment | Organism | Inhibitor | Structure |
---|---|---|---|---|---|
0.0252 | - |
pH 8.0, 25°C, recombinant enzyme | Plasmodium falciparum | bithionol | |
0.12 | - |
pH 8.0, 25°C, recombinant enzyme | Plasmodium falciparum | bithionol | |
0.139 | - |
pH 8.0, 25°C, recombinant enzyme | Plasmodium falciparum | GW5074 | |
0.15 | - |
pH 8.0, 25°C, recombinant enzyme | Plasmodium falciparum | GW5074 |
General Information | Comment | Organism |
---|---|---|
additional information | the parasitic enzyme does not contain the antenna domain, responsible for allosteric regulation in the mammalian enzymes | Plasmodium falciparum |
additional information | the three-dimensional structure of hexameric PfGDH2 is solved to 3.1 A resolution, overview. The parasitic enzyme does not contain the antenna domain, responsible for allosteric regulation in the mammalian enzymes | Plasmodium falciparum |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.75 | - |
L-glutamate | pH 8.0, 25°C, recombinant enzyme | Plasmodium falciparum | |
6 | - |
L-glutamate | pH 8.0, 25°C, recombinant enzyme | Plasmodium falciparum | |
14.67 | - |
2-oxoglutarate | pH 7.0, 25°C, recombinant enzyme | Plasmodium falciparum | |
1250 | - |
2-oxoglutarate | pH 7.0, 25°C, recombinant enzyme | Plasmodium falciparum |